Animal Tissue Culture Basics
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Questions and Answers

Which of the following substances is classified as a carcinogen?

  • Streptomycin
  • Hygromycin (correct)
  • Gentamycin
  • Thapsigargin

What is cellular senescence?

  • The process of cells undergoing mutation
  • The phenomenon where cells lose the ability to divide (correct)
  • The ability of cells to divide indefinitely
  • The method of isolating cells from tissues

What method is used to release mononuclear cells from soft tissues for culture?

  • Mechanical disruption of tissue
  • Using a centrifuge to separate components
  • Placing tissue pieces in saline solution
  • Enzymatic digestion with collagenase (correct)

Which of the following statements about primary cell cultures is accurate?

<p>They have a limited lifespan with a defined number of population doublings. (A)</p> Signup and view all the answers

What defines an established or immortalized cell line?

<p>Cells that can proliferate indefinitely. (B)</p> Signup and view all the answers

What is a primary cell culture primarily derived from?

<p>Dispersed cells from the original tissue (A)</p> Signup and view all the answers

What characterizes an established or continuous cell line?

<p>It has infinite growth potential due to transformation. (C)</p> Signup and view all the answers

What is the purpose of a feeder layer in cell culture?

<p>To serve as a substrate for other cells to grow on. (B)</p> Signup and view all the answers

Which of the following is a common application of tissue culture?

<p>Producing monoclonal antibodies. (C)</p> Signup and view all the answers

What type of substrate may be used in cell culture?

<p>Solid, semisolid, or liquid (A)</p> Signup and view all the answers

What does passage number refer to in cell culture?

<p>The number of times cells have been transferred to a new vessel. (A)</p> Signup and view all the answers

Which factor is NOT typically regulated in the culture environment of cells?

<p>Color of the culture medium (C)</p> Signup and view all the answers

What is one feature of the media choice in cell culture?

<p>It is determined empirically for specific cell types. (A)</p> Signup and view all the answers

What is the primary purpose of cell/tissue culture?

<p>To propagate cells outside the organism (D)</p> Signup and view all the answers

Which of the following statements is true about tissue culture?

<p>Tissue culture results in clones with the same genotype (C)</p> Signup and view all the answers

What significant advancement did Enders, Weller, and Robbins achieve in 1948?

<p>Proved poliomyelitis virus can be grown in vitro without nerve tissue (D)</p> Signup and view all the answers

What was the main contribution of George Gey in 1952?

<p>Establishing the first human cell line from cervical carcinoma (C)</p> Signup and view all the answers

What is the primary challenge associated with growing large complex proteins in E. coli?

<p>E. coli does not provide proper glycosylation (D)</p> Signup and view all the answers

Which technique allows for the maintenance of the structural integrity of tissue in culture?

<p>Organ culture (A)</p> Signup and view all the answers

Which of the following describes a significant aspect of the 1955 advances in cell culture media?

<p>Provided consistency and reduced chances of contamination (C)</p> Signup and view all the answers

What is a characteristic of normal human diploid cells as described by Hayflick and Moorhead?

<p>They exhibit a finite life span (C)</p> Signup and view all the answers

What is a major difference between transformed cells and primary cells in cell culture?

<p>Transformed cells can grow without cell-cell contact. (C)</p> Signup and view all the answers

Which of the following is NOT a limitation of cell culture?

<p>Total inability to grow any cells (C)</p> Signup and view all the answers

What is a significant feature that cell culture lacks when compared to normal tissue?

<p>Blood circulation (C)</p> Signup and view all the answers

Which characteristic of transformed cells can lead to variability in experimental outcomes?

<p>High adaptability to various conditions (C)</p> Signup and view all the answers

What might be a consequence of using hormones in high concentrations in cell culture?

<p>Alteration in cell types (D)</p> Signup and view all the answers

Which factor contributes to the genetic instability of cells in culture?

<p>Heterogeneity of cell populations (B)</p> Signup and view all the answers

What is a critical environmental control issue when conducting cell culture experiments?

<p>Maintaining a specific pH level (C)</p> Signup and view all the answers

Why is cell contact significant for primary cells in culture?

<p>It affects genetic expression. (D)</p> Signup and view all the answers

What characteristic of tissue culture allows for experiments to be conducted without the variations seen in animal studies?

<p>Control of the growth environment (D)</p> Signup and view all the answers

Which of the following advantages is a characteristic of tissue culture?

<p>Ability to expose cultures to defined concentrations of reagents (B)</p> Signup and view all the answers

One of the limitations of tissue culture is that it lacks the physiological complexity of living organisms. What specifically is this limitation related to?

<p>Absence of organized tissues (D)</p> Signup and view all the answers

How can tissue cultures aid in reducing ethical concerns related to research?

<p>By minimizing animal experimentation (D)</p> Signup and view all the answers

What is a common disadvantage of tissue culture that researchers must address?

<p>Dedifferentiation and potential loss of cellular mechanisms (B)</p> Signup and view all the answers

Which factor can be controlled in the physico-chemical environment of tissue culture?

<p>Osmolarity and pH levels (A)</p> Signup and view all the answers

Which aspect of tissue culture helps ensure that a consistent lineage of cells is maintained over generations?

<p>Selective media and cloning availability (D)</p> Signup and view all the answers

What is a characteristic challenge faced by researchers when working with tissue cultures?

<p>Learning and maintaining aseptic techniques (A)</p> Signup and view all the answers

Flashcards

Cell Culture

The process of growing cells outside of an organism, including both plant and animal cells.

Cloning in Cell Culture

When cells are grown in a lab dish and they divide, making copies of themselves with the same genetic material. This is like taking a single plant and making many identical copies.

Environmental Conditions for Cell Culture

The conditions in which cells are grown in a lab dish must be very similar to the conditions that the cells normally experience in the organism. This includes temperature, humidity, food, and keeping things clean.

Organ Culture

A type of cell culture where a whole piece of tissue, with its 3D structure, is kept alive in a dish. This is like keeping a small piece of an organ working in the lab.

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Cell Culture

Growing single cells in a dish, after they've been taken apart from the tissue they were originally in, and then grown in the lab.

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Primary Cell Culture

When cells are directly taken from an organism and put into a dish to grow. This is like taking a fresh piece of fruit and growing it in a greenhouse.

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Clone

A group of cells that are all originally from a single cell, like identical twins. These cells are all genetically identical.

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Subculture

Transferring cells from one container to a new container to give them more space to grow, like moving plants to a larger pot.

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Established Cell Lines

Cell cultures that can keep dividing and growing forever. These are often cells that come from tumors.

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Passage Number

The number of times that cells have been subcultured or moved to new containers. Like keeping track of how many times a plant has been repotted.

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Research Applications of Cell Culture

Using cell cultures to study how cells work and how chemicals affect them. It's like conducting experiments on cells to see how they behave.

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Tissue Engineering Applications of Cell Culture

Growing cells in a lab to create tissues like skin, liver, bone, and cartilage. This is like building a new organ from scratch.

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Production of Biologicals using Cell Culture

Using cell cultures to produce important substances like antibodies, hormones, and vaccines. This is like using cells as little factories to produce valuable medicines.

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Substrates for Cell Culture

Materials that cells can grow on in a dish, which can be solid, like glass or plastic, semi-solid, like jelly, or liquid, allowing cells to float freely.

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Feeder Layers in Cell Culture

Cells that help other cells grow by providing them with extra support. They act like friendly neighbors in the cell world.

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Gas Phase for Cell Cultures

The air that cells breathe in a lab dish must have the right mixture of carbon dioxide and oxygen for them to thrive.

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Media for Cell Cultures

The liquid that gives cells food and everything they need to grow in a dish. It's like a cell's special drink and meal.

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Advantages of Cell Culture: Studying Individual Cells

The benefit of being able to study cells individually and see how they behave without interference from other cells and tissues in the organism.

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Advantages of Cell Culture: Controlled Environment

Cell cultures can be grown in a highly controlled environment, so all the cells are very similar to each other, making the results of experiments more reliable.

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Advantages of Cell Culture: Reproducibility

Cell cultures can be used to study how cells change over time and still keep the same characteristics, so scientists can repeat experiments and get the same results.

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Advantages of Cell Culture: Scaling Up

Cell cultures can be used to produce large numbers of cells, like growing lots of crops, which is useful for research and for making medicines.

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Advantages of Cell Culture: Ethics

Cell cultures can be used to study cells and tissues without directly using animals. This is a more ethical way to do research.

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Shortcomings of Cell Culture: Limited Conditions

The disadvantage of cell cultures that they lack the full complexity and natural conditions of how cells behave in a living organism.

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Shortcomings of Cell Culture: Dedifferentiation

Cells in a dish can sometimes lose their specific characteristics and start to behave like more basic cells. It's like a cell forgetting its identity.

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Shortcomings of Cell Culture: Standardized Techniques

The techniques for growing cells in culture must be very precise to ensure cells grow properly and give consistent results.

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Shortcomings of Cell Culture: Costs

The expense of materials and the cost of maintaining cell cultures can limit the amount of research that can be done.

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Missing Features of Cell Culture

Cells growing in a dish miss features like blood circulation, how tissues organize, and how cells interact with each other, things that are essential in a living organism.

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Missing Features of Cell Culture: Hormones

In cell cultures, hormones need to be added in high concentrations, unlike in the body where they are present in small amounts naturally.

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Safety in Cell Culture

The need to be careful when working with cell cultures as they can contain substances that are dangerous if handled improperly.

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Types of Cell Culture: Isolating Cells

Cells can be taken from tissues using techniques like enzymatic digestion to break down the tissue or by carefully growing a small piece of tissue in a dish.

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Types of Cell Culture: Primary Cells

Primary cells are directly taken from an organism and have a limited number of times they can divide before they stop growing. It's like a cell lifespan.

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Types of Cell Culture: Established Cell Lines

Established cell lines are cells that can keep dividing and growing indefinitely, often derived from tumors. They're cells with a long lifespan, like a long-lasting cell line.

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Study Notes

Overview of Cell/Tissue Culture

  • Cell culture is the propagation of cells outside an organism, involving both plant and animal cells.
  • Tissue culture produces clones with identical genotypes unless mutations occur during culture.
  • Environmental conditions in culture must replicate the organism's conditions: temperature, humidity, nutrition, and contamination-free.

History of Cell Culture

  • 1801: Bichat names human body tissues.
  • 1880: Roux maintains embryonic chicken tissue in saline.
  • 1907: Harrison demonstrates in vitro growth of living animal tissue from a tadpole nerve cord.
  • 1911-1912: Burrows and Carrel grow explants from adult animals, employing aseptic techniques.
  • 1948: Enders, Weller, and Robbins grow polio virus in vitro using monkey kidney cells.
  • 1952: George Gey establishes the HeLa cell line from human cervical carcinoma.
  • 1961: Hayflick and Moorhead identify finite lifespan of normal human diploid cells.

Terminology

  • Organ culture: Three-dimensional culture retaining tissue features in vivo.
  • Cell culture: Single cells derived from dispersed tissue cells.
  • Primary cell culture: Directly derived from explants with a limited lifespan.
  • Clone: A population derived from a single original cell.
  • Sub-culture: Transplantation of cells to new vessels.
  • Established cell lines: Immortalized cell cultures, often tumor-derived, like CHO and SH-SY-5Y cells.
  • Passage number: Successive sub-cultures from the primary culture.

Applications of Cell Culture

  • Research: Understand cell functions and toxicology.
  • Tissue engineering: Cultivation of skin, liver, bone, and cartilage.
  • Production of biologicals: Create monoclonal antibodies, hormones, enzymes, and vaccines.

Culture Environment

  • Substrates can be solid (glass, plastic), semisolid (agar, collagen gel), or liquid (suspension cultures).
  • Feeder layers support growth by preventing proliferation.
  • Gas phase: Standard CO2 and O2 levels must be maintained to optimize cell growth.
  • Media must provide nutrients and be cell-type specific, ensuring sterility and isotonic conditions.

Advantages of Tissue Culture

  • Enables cell behaviour study without variations from whole organisms.
  • Controlled growth environments ensure sample uniformity.
  • Cells retain characteristics over generations, providing reproducibility.
  • Large quantities of cells can be produced.
  • Reduces ethical concerns related to animal experimentation.

Shortcomings and Limitations

  • In vitro culture lacks true physiological conditions and the complex interactions present in vivo.
  • Cells can undergo dedifferentiation, leading to loss of original features.
  • Standardized techniques are required for successful cell maintenance.
  • Limited material quantity and costly consumables can be a barrier.

Missing Features in Cell Culture

  • Unlike in vivo conditions, blood circulation, tissue organization, and cellular interaction are often absent in cultures.
  • Hormones are typically added in high concentrations, unlike natural physiological levels.

Safety in Cell Culture

  • Be aware of hazardous substances: carcinogens, teratogens, and mutagens.
  • Common examples include gentamycin (possible teratogen), hygromycin (possible carcinogen), and streptomycin (mutagen).

Types of Tissue Culture

  • Cells isolated from tissues can be cultured using enzymatic digestion techniques or explant cultures.
  • Primary cells have a limited lifespan, undergoing senescence after several divisions.
  • Established or immortalized cell lines can proliferate indefinitely, often used for research and diagnostics.

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Description

Explore the fundamentals of animal tissue culture, a technique for growing and maintaining cells outside their natural environment. This quiz covers the principles of cell and tissue culture, including the propagation of genetically identical clones. Test your knowledge on the methods and applications of this crucial biological technique.

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